pE-4000

The pE-4000 sets the standard as the universal Illumination System for fluorescence microscopy.

Overview

The pE-4000 sets the standard as the universal Illumination System for fluorescence microscopy. The system has 16 selectable LED wavelengths across four channels that can be finely controlled and matched to the filters and fluorophores of almost any microscope, making it the broadest and most versatile illumination system available.

The CoolLED pE-4000 benefits from our award winning sustainable Green technology and delivers enhanced intensity at the sample plane with a significant reduction in energy consumption, and is supplied with a 36 month warranty.

The pE-4000 is a powerful, flexible, controllable system for advanced research applications.

“Striking, bright fluorescence images with a strong signal to noise ratio even when using a low magnification objective”

Graham Wright, Institute of Medical Biology, A*STAR Singapore

Advanced control for cutting-edge research

Multiple levels of control are possible, from simple to advanced. A dual mode control pod gives immediate manual control in either White or Advanced modes, where individual LED channels can be individually switched on/off or intensity modulated.

Alternatively, the USB interface and integration into major microscope software platforms allows complex software control.
TTL control enables capturing high-speed events (<1 ms). Since there is no shutter or filter wheel, without these moving parts, this reduced latency means speed is limited only by the camera. An expansion box is also available for added functionality with multiple BNC connections for remote analogue and TTL control.

For optogenetics applications, alongside the ability to include excitation filters to tune the wavelength to the opsin’s maximum activation peak, an internal function generator controls pulse timing and waveforms.

Improving cell viability

With the pE-4000, it is now possible to prevent artefacts and allow live samples to thrive in experiments >100 hours. Fine control of illumination timing and intensity with TTL and the function generator minimises exposure, avoiding photobleaching and phototoxicity.

“When you can only control the intensity of ‘white’ light (rather than individual channels), the level of photobleaching can be high. With the pE-4000, we can control the excitation of the individual channels. It is possible to optimise the excitation intensity according to the labelling, greatly reducing photobleaching and phototoxicity in a live experiment.”

Dr Yan Gu, University of Sussex

“Suddenly, we were able to offer users uninterrupted extended live cell experiments of 100+ hours, without worrying about brightness fluctuations, lamps burning, room heating, etc. Also, users have reported markedly reduced bleaching and phototoxicity in their samples, both from the prokaryotic and the eukaryotic research fields.”

Dr Jens Eriksson, Oslo University Hospital

Matching diverse fluorophore requirements

Full filter compatibility and 256 wavelength combinations from 16 installed LEDs (365 nm -770 nm) covers all available fluorophores, which is ideal for multi-user facilities or evolving experimental needs.

Multi-channel experiments with no pixel shift

Up to four channels using distinct wavelengths can be captured with no mechanical movement, preventing pixel shift and allowing the direct comparison of data as in the following experiment:

(A) Representative photocurrent traces of a PhobosCA expressing CA1 cell evoked with different activation wavelengths and shutoff with 595 nm light. (B) Photocurrent traces in the same cell evoked with 460 nm light and shutoff with indicated wavelengths (10 mW/mm2).1

Increased power efficiency

Our unique Wavelength Grouping Concept makes it possible to deliver more power in an efficient four-channel system. As the first company to introduce LEDs into fluorescence microscopy, CoolLED has developed a comprehensive understanding of the complexities of configuring and selecting filter sets for experiments using multiple fluorophores. CoolLED’s innovation comes from recognising that all the fluorophores used in multi-band work can have their absorption bands divided into four separate groups across the spectrum due to the restricted bandwidth available.

Choose liquid light guide or fibre

The pE-4000 can be configured to deliver light to the microscope via liquid light guide or fibre. Liquid light guides can be used in conjunction with CoolLED’s pE-Universal Collimator which can accept a microscope adaptor from CoolLED’s extensive range. Find out more here.

I have been using a pE-4000 for optogenetic experiments using in vitro slices for just over a year now. The pE-4000 has been fantastic for my needs: the ability to simultaneously use multiple wavelengths of light has allowed me to carry out experiments using two different opsins to study how external inputs converge on hippocampal interneurons. I’m currently setting up my own research group at the University of Exeter, and all of my patch-clamp rigs will have a pE-4000 on them, as they are such versatile devices for optogenetic experiments.

Dr Michael Craig, Research Fellow, University of Exeter Medical School

Our group is currently attempting to isolate and stain circulating tumour cells. In order to identify these cells we need a minimum of four colour fluorescence imaging, requiring the slide to be scanned using four different LEDs.To stitch our images together after scanning we require a transmission light microscope image. The image co-ordinates of the light microscope image are used to stitch the fluorescence images together. The previous light source (PE-2) allowed only three LEDs to be used together for our purposes. This meant that when we have multiple slides there are two options to get the full four fluorescence scan:1) Change the LED after scanning in Brightfield and three fluorescence channels – this risks damage to the LED itself and is time consuming. It involves un-screwing a panel each time and screws can be damaged or lost relatively easily. It places demand on the ribbon that is connected to the LED and, over time, may damage the ribbon also. We usually scan multiple slides and prefer to use option two, to reduce wear and tear on the light source.2) This option is to scan all slides in Brightfield and three colour fluorescence and then go for a second batch of images in both Brightfield and the final fluorescence channel. This is also time consuming, as setting up each scan in focus takes time. It also requires a lot more downstream processing to line the images up with one another, as the scans are often of two different sizes.With the new PE-4000 light source I was able to scan four slides in Brightfield and four fluorescence channels in three hours. Four slides took six hours to scan on the previous light source, due to double the number of scans and changing the LED. It has also reduced the downstream processing of the images, as they are all taken using the same co-ordinates, in one batch.Our project aims to expand from four colour fluorescence to six or seven channels over the coming years. Using the old light sources this would require a minimum of three but possibly four scans taken from different positions per slide and would take hours per slide.The current light source meets the needs of researches using modern immunofluorescence techniques as most projects have moved from single to a multi-marker approach, in recent years.

I have been imaging using the new CoolLED PE-4000 system on our epi-fluorescence microscope. This is an excellent system as it covers a very wide spectrum of wavelengths. I have found it very easy to use. The user interface is much better as the different LEDs can be changed using the handheld control- without switching off and unscrewing the LAMs as had to be done on the old PE-2 system. In addition the LEDs can be changed dynamically while the software to capture the image is open, and also a huge combination of different wavelengths can be used at the same time.I have found that the light source from this system is considerably stronger than from the previous system - I now image at 50% strength, when I used to image at 100%.I anticipate using this imaging system a lot more now that the new LED light source is there.

I am very impressed with the brightness and performance of the CoolLED pE-4000 in comparison to the mercury or halide light sources for fluorescence. I also really like its neat packaging and it is easy to set up and configure. Absolutely brilliant upgrade for archaic mercury set ups!

Dominic White, Area Sales Manager, Carl Zeiss Microscopy GmbH

Throwing out the old mercury lamp and exchanging it for the pE-4000 on one of our live cell wide field microscopes has been an astounding success! It allowed us to cheaply breathe new life into old equipment. Suddenly, we were able to offer users uninterrupted extended live cell experiments of 100+ hours, without worrying about brightness fluctuations, lamps burning, room heating, etc. Also, users have reported markedly reduced bleaching and phototoxicity in their samples, both from the prokaryotic and the eukaryotic research fields. As a facility manager I am pleased with the instrument's ease of use, and shallow learning curve for new users. Taken together with reduced overall microscope maintenance, the pE-4000 has already saved me a substantial amount of time in my daily work life, and improved the quality of our services at the same time.

Conceptually, the pE-4000 seems an extremely versatile tool that blends seamlessly with our range of microscopes, making it ideal for high end life science applications. It is also very user friendly and powerful.

Dr Alex Gardiol, PhD, Imaging Specialist, Olympus Microscopy

I just purchased a pE-4000 to replace an old version of Lumencor Sola SE on a live imaging microscope. After comparing the power level of the systems, I am convinced that the pE-4000 provides stronger illumination at the objective. As the Sola light source can only control intensity of ‘white’ light (rather than individual channels), we have to choose the excitation level according to the dimmest labelling fluorophore. The other channels are in turn exposed to an unnecessary high dose of excitation. The level of photo-bleaching can be high as a result. With the pE-4000, we can control the excitation of the individual channel. It is possible to optimise the excitation intensity according to the labelling, greatly reducing photo-bleaching and photo-toxicity in a live experiment. Due to the difference in the excitation spectrum with the Sola, I was worried about the extra cost of changing the dichroic, excitation and emission filters. The options of four wavelength in each channel (totally four channels) enables me to pick up the correct wavelength according to the old filter/dichroic settings.

A keratinocyte undergoing mitosis in which the keratin is labelled with GFP. Acquired on a Olympus IX-83 microscope – CoolLED pE-4000. Scale bar = 10 μm.

CoolLED pE-4000 working with cultured hippocampal neurons

Fluorescence image of 15 zebrafish larvae (fixed and stained with acridine orange) within the field of the Mesolens.

Time series of a confluent sheet of keratinocytes migrating during a live-cell imaging outgrowth assay, acquired on an Olympus IX-83 microscope equipped with a CoolLED pE-4000 lightsource (Fig. 4). Scale bar = 150 μm.

Fibres

Liquid Light Guide

Interchangeable Light Delivery Output module for LLG (3 mm diameter)

Filter Holders

pE-6501

The CoolLED pE Driver is first required for software integration, and can be downloaded here.

Company

Software

Summary

Screenshot

Quick Start Guide

Andor

Andor iQ

Hamamatsu

HCImage

Intelligent Imaging Innovations (3i)

Slidebook

LaVision BioTec

ImSpector Pro

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Leica

LASX

Micromanager

Micromanager

Molecular Devices

MetaMorph

Nikon

NIS Elements

Olympus

CellSens

Visitron Systems

Visiview

&nbsp

Zeiss

ZEN

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CoolLED Proprietary ZEN Bridge software is available – Contact CoolLED for further information.

Environment & Safety

Did you know that LED illumination systems help laboratories reduce their carbon footprint, protecting laboratory staff along the way?

Toxic mercury-based microscope illuminators are bad news for the environment, draw a lot of power and have a short lifetime.

LED illumination systems are cleaner, have a long lifetime and use less power. Our pE-300 Series is also ACT label certified and is a natural choice for labs who want to play their part in helping the environment.

Help & Advice

At CoolLED we are committed to giving our customers outstanding service right from the start and throughout the lifetime of the product. Please get in touch if you have any questions or would like a quote. You can also browse our site for technical resources and general support.